US9062126B2ExpiredUtilityPatentIndex 81

Compositions comprising receptor-associated protein (RAP) variants specific for CR-containing proteins and uses thereof

Assignee: ZANKEL TODD CPriority: Sep 16, 2005Filed: Sep 18, 2006Granted: Jun 23, 2015

Est. expirySep 16, 2025(expired)· nominal 20-yr term from priority

Inventors:ZANKEL TODD C STARR CHRISTOPHER M

A61P 5/00A61P 37/06A61P 3/06A61P 9/00A61P 25/18A61P 25/22A61P 25/16A61P 35/00A61P 25/28A61P 25/24A61P 25/00A61P 25/08A61P 31/04A61P 21/02C07K 14/705A61K 38/00A61K 38/1709A61K 47/64A61P 21/00C07K 14/47A61K 47/48246

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Cited by

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References

Claims

Abstract

The present invention relates generally to receptor-selective variants of the low-density lipoprotein receptor-associated protein (RAP) and compositions thereof, methods of generating such variants and methods of using such receptor-selective RAP variant compositions for therapeutic purposes.

Claims

exact text as granted — not AI-modified

What is claimed is: 
     
       1. An isolated polypeptide comprising an amino acid sequence variant of the mature alpha-2-macroglobulin/low density lipoprotein receptor-related protein-associated protein 1 (RAP) set out in SEQ ID NO: 121, wherein said RAP variant binds selectively to a pair of complement-type repeats within a protein selected from the group consisting of very low density lipoprotein receptor (VLDLR), matriptase and CD320, with an affinity that is at least 3-fold greater than the affinity of wild-type RAP for the same pair of complement-type repeats, wherein the RAP variant consists of
 (a) a polypeptide having an amino acid sequence at least 90% identical to SEQ ID NO: 121 and comprises mutations at at least two of the following positions: 251, 256, 257, 266 and 270, and optionally comprises a mutation at one or more of positions 175, 205, 213, 217, 226, 230, 232, 239, 246, 249, 251, 256, 257, 261, 266, 267, 268, 270, 273, 287, 290, 294, 296, 297, 298, 305, 312 or 313 of mature RAP; or 
 (b) a polypeptide having an amino acid sequence at least 90% identical to amino acids 200-323 of SEQ ID NO: 121 and comprises mutations at at least two of the following positions: 251, 256, 257, 266 and 270, and optionally comprises a mutation at one or more of positions 205, 213, 217, 226, 230, 232, 239, 246, 249, 251, 256, 257, 261, 266, 267, 268, 270, 273, 287, 290, 294, 296, 297, 298, 305, 312, or 313 of mature RAP. 
 
     
     
       2. The polypeptide of  claim 1  wherein said RAP variant lacks amino acids 1-143 and 320-323 of a mature RAP. 
     
     
       3. A RAP variant of  claim 1  missing at least 200 and up to 243 amino acids from the N-terminus. 
     
     
       4. The RAP variant of  claim 3  further missing up to 11 amino acids from the C-terminus. 
     
     
       5. The RAP variant of  claim 4  missing at least 4 amino acids from the C-terminus. 
     
     
       6. The RAP variant of  claim 1  comprising a continuous portion of mature RAP that is (a) at least 71 amino acids in length and (b) comprises amino acids 256-270. 
     
     
       7. The RAP variant of  claim 1  comprising a continuous portion of RAP d3 that is (a) at least 71 amino acids in length and (b) comprises amino acids 256-270. 
     
     
       8. The polypeptide of  claim 7  wherein the variant contains a mutation at any one of positions 251, 256, 257, 266, 270, 279, 280, 296 or 305 of RAP. 
     
     
       9. The polypeptide of  claim 1  wherein the variant binds selectively to a matriptase protein. 
     
     
       10. The polypeptide of  claim 9  wherein the variant contains a mutation at any one of positions 251, 256, 257, 266, 270, or 280 of RAP. 
     
     
       11. The polypeptide of any one of  claims 9 - 10  wherein the variant comprises a mutation at position 251 of RAP. 
     
     
       12. The polypeptide of any one of  claims 9 - 10  wherein the variant comprises a mutation at position 256 of RAP. 
     
     
       13. The polypeptide of any one of  claims 9 - 10  wherein the variant comprises a mutation at position 14 of RAP. 
     
     
       14. The polypeptide of any one of  claims 9 - 10  wherein the variant comprises a mutation at position 266 of RAP. 
     
     
       15. The polypeptide of any one of  claims 9 - 10  wherein the variant comprises a mutation at position 270 of RAP. 
     
     
       16. The polypeptide of any one of  claims 9 - 10  wherein the variant comprises a mutation at position 280 of RAP. 
     
     
       17. The polypeptide of  claim 1  wherein the variant binds selectively to a VLDLR protein. 
     
     
       18. The polypeptide of  claim 17  wherein the variant contains mutations at at least two positions selected from any one of positions 251, 256, 270 or 296 of RAP. 
     
     
       19. The polypeptide of any one of  claims 17 - 18  wherein the variant comprises a mutation at position 251 of RAP. 
     
     
       20. The polypeptide of any one of  claims 17 - 18  wherein the variant comprises a mutation at position 256 of RAP. 
     
     
       21. The polypeptide of any one of  claims 17 - 18  wherein the variant comprises a mutation at position 270 of RAP. 
     
     
       22. The polypeptide of any one of  claims 17 - 18  wherein the variant comprises a mutation at position 296 of RAP. 
     
     
       23. The polypeptide of  claim 1  wherein the variant binds selectively to a CD320 protein. 
     
     
       24. The polypeptide of  claim 23  wherein the variant contains a mutation at any one of positions 251, 256, 270, 279 or 305 of RAP. 
     
     
       25. The polypeptide of any one of  claims 23 - 24  wherein the variant comprises a mutation at position 251 of RAP. 
     
     
       26. The polypeptide of any one of  claims 23 - 24  wherein the variant comprises a mutation at position 256 of RAP. 
     
     
       27. The polypeptide of any one of  claims 23 - 24  wherein the variant comprises a mutation at position 270 of RAP. 
     
     
       28. The polypeptide of any one of  claims 23 - 24  wherein the variant comprises a mutation at position 279 of RAP. 
     
     
       29. The polypeptide of any one of  claims 23 - 24  wherein the variant comprises a mutation at position 305 of RAP. 
     
     
       30. The polypeptide of  claim 1  wherein the variant contains at least one additional mutation within positions 271-319 of RAP. 
     
     
       31. The polypeptide of  claim 1  wherein the variant contains at least one mutation within positions 205-250 of RAP. 
     
     
       32. The polypeptide according to any one of  claims 8 ,  9 ,  17 , or  23 , wherein any of the mutations comprises the replacement of an acidic amino acid with a basic amino acid. 
     
     
       33. The polypeptide of  claim 32 , wherein said acidic amino acid is selected from the group consisting of D and E. 
     
     
       34. A polypeptide of  claim 32 , wherein said basic amino acid is selected from the group consisting of K and R. 
     
     
       35. A polypeptide according to any one of  claims 8 ,  9 ,  17 , or  23 , wherein any of the mutations comprises the replacement of a basic amino acid with an acidic amino acid. 
     
     
       36. A polypeptide of  claim 35 , wherein said basic amino acid is selected from the group consisting of K and R. 
     
     
       37. A polypeptide of  claim 35 , wherein said acidic amino acid is selected from the group consisting of D and E. 
     
     
       38. A polypeptide according to any one of  claims 8 ,  9 ,  17 , or  23 , wherein any of the mutations comprises the replacement of an amino acid selected from the group consisting of A, C, D, E, G, I, K, L, M, N, P, Q, R, S, T, and V with an amino acid selected from the group consisting of F, Y, W, and H. 
     
     
       39. The polypeptide of any one of  claims 1 ,  7 ,  8 ,  9 ,  17 , or  23  that comprises a mutation at three or more of the following positions: 175, 205, 213, 217, 226, 230, 232, 239, 246, 249, 251, 256, 257, 261, 266, 267, 268, 270, 273, 287, 290, 294, 296, 297, 298, 305, 312, 313. 
     
     
       40. The polypeptide of any one of  claims 1 ,  7 ,  8 ,  9 ,  17 , or  23  that comprises a mutation at three or more of the following positions: 205, 217, 249, 251, 256, 257, 266, 270, 294, 296, 297, 305. 
     
     
       41. The polypeptide of any one of  claims 1 ,  7 ,  8 ,  9 ,  17 , or  23  that comprises three or more of the following mutations: R205S, S213T, E217K, L226M, H249Y, A230V, S232P, E239G, E246G, E251L, E251K, E251T, E251G, E251P, E251N, E251R, K256R, K256V, K256A, K256I, K256P, K256L, I266F, I266T, H257Y, Q261R, A267V, H268R, K270P, K270D, K270N, K270G, K270E, K270W, L271M, H273Y, D279Y, V283M, R287H, H290Y, H290L, E294V, R296L, T297I, K298R, K305T, K306M, S312F, G313D. 
     
     
       42. A polypeptide consisting of oligomeric combinations of polypeptides of any one of  claims 1 ,  7 ,  8 ,  9 ,  17 , or  23  with binding multivalent binding properties defined by each polypeptide of the combination. 
     
     
       43. A compound comprising the polypeptide according to any one of  claims 1 ,  7 ,  8 ,  9 ,  17 , or  23  conjugated to a diagnostic or therapeutic agent. 
     
     
       44. The compound of  claim 43 , wherein the polypeptide and diagnostic or therapeutic agent are linked through a linker. 
     
     
       45. The compound of  claim 44 , wherein said linker is a peptide linker. 
     
     
       46. The compound of  claim 43 , wherein the therapeutic agent is selected from the group consisting of a glial cell-derived neuronal growth factor (GDNF), brain-derived neuronal growth factor (BDNF), neuronal growth factor (NGF), disintegrin and metalloproteinase domain 10 (ADAM10), a chaperone protein for LRPS/6 (MESD), cancer chemotherapeutic agents, protease inhibitors, pro-apoptotic molecules, autoimmune antigens, lysosomal enzymes, nanoparticles, glycoconjugates and nucleic acids. 
     
     
       47. A pharmaceutical composition comprising a compound of  claim 43  in a pharmaceutically acceptable carrier, diluent or excipient.

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